Journal: Critical Care

Article Title: Endothelial permeability following coronary artery bypass grafting: an observational study on the possible role of angiopoietin imbalance

doi: 10.1186/s13054-016-1238-0

Figure Lengend Snippet: Influence of angiopoietin ( ANG )2/ANG1 imbalance on endothelial hyper-permeability ex vivo . Human pulmonary microvascular endothelial cells were seeded at high density on gelatin-coated permeable supports and cultured to form a confluent monolayer. After pre-incubation with filtered and diluted (7.5 %) patient serum (preoperative ( preop ) or 24 h, respectively) for 30 minutes, fluorescein isothiocyanate ( FITC )-conjugated dextran (70 kDa, final concentration 1 mg/ml) was added to the upper compartment and the inserts were set on a magnetic stirrer. Thrombin (final concentration 1 IU/ml) was added to induce endothelial permeability. Samples (of 50 μl) were taken from the lower compartment at the indicated time points. Dextran concentration was determined using a fluorescent plate reader. Thrombin-induced hyper-permeability is expressed as fold of the average basal flux during the initial 15-minute equilibration period before thrombin addition. Left , change in endothelial cell (EC) permeability after pre-incubation with serum samples with a mild shift in the ANG2/ANG1 ratio upon coronary artery bypass graft (CABG) (6-fold from baseline to 24 h). Right , change of EC permeability after pre-incubation with serum samples with pronounced shift in the ANG2/ANG1 ratio upon CABG (18-fold from baseline to 24 h). N = 8–10, mean ± standard error of the mean, * p <0.05, ** p <0.01. Pat patient

Article Snippet: Fluorescence was measured using a fluorescent plate reader (excitation 492 nm, emission 518 nm; Modulus II Microplate Multimode Reader, Promega, Fitchburg, WI, USA).

Techniques: Permeability, Ex Vivo, Cell Culture, Incubation, Concentration Assay